Development of real‐time detection direct test for hepatitis B virus and comparison with two commercial tests using the WHO international standard

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<jats:title>Abstract</jats:title><jats:p><jats:bold>Aims: </jats:bold> A highly reproducible and sensitive hepatitis B virus real‐time detection direct (HBV RTD‐direct) test using DNA extraction by magnetic beads coated with polyclonal anti‐HBsAg, followed by the real‐time detection polymerase chain reaction (PCR) method, was developed for the detection of HBV DNA.</jats:p><jats:p><jats:bold>Methods: </jats:bold> The HBV DNA could be extracted from the HBsAg positive viral particles without resulting in viral DNA fragmentation. The HBV RTD‐direct test was validated using a serial dilution panel of the WHO standard HBV DNA 97/746 I.</jats:p><jats:p><jats:bold>Results: </jats:bold> The test had a dynamic range of 0.7–8.0 log<jats:sub>10</jats:sub> international units (IU) per mL and the results were shown to be comparable to those obtained with two commercially available tests: the HBV DNA transcription‐mediated amplification‐hybridization protection assay and the Amplicor HBV Monitor test. In addition, the HBV RTD‐direct test, based on magnetic extraction, successfully eliminated PCR inhibitors in clinical specimens.</jats:p><jats:p><jats:bold>Conclusion: </jats:bold> We conclude that the HBV RTD‐direct test is an excellent alternative for monitoring patients undergoing antiviral treatment or for screening various clinical specimens.</jats:p>

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